Restriction digests of the clone give the following sizes (kb): BamHI--33.0, 9.4; EcoRI--33.0, 9.4; BglII--22.0, 8.8, 4.8, 4.6, 3.1; PstI--9.6, 9.0, 4.6, 3.2, 2.9, 2.8, 2.4, 2.2, 1.9, 1.9, 1.5. Vector allowing expression of cloned inserts as a fusion protein on the phage particle surface. Inserts are fused to the C-terminus of a truncated phage tail protein by a peptide linker. Presence of the lacZalpha coding sequence and ribosome binding site allow blue-white color detection of recombinants, as well as allowing expression of a cloned insert separate from the phage tail protein. The Pro-Thr box encodes alternating prolyl and threonyl residues, which form a link between the N-terminal phage tail protein and the foreign protein. The linker resembles the hinge-region of IgA1 and allows separation of the foreign protein from the phage particles by digestion with enzymes such as Cellulomonas fimi protease or collagenase. Production of large amounts of fusion protein may inhibit phage assembly. A host allowing low efficiency suppression of the amber mutation is recommended. |
